Non-targeted and targeted analysis of oxylipins in combination with charge-switch derivatization by ion mobility high-resolution mass spectrometry

Abstract

Eicosanoids and other oxylipins play an important role in mediating inflammation as well as other biological processes. For the investigation of their biological role(s), comprehensive analytical methods are necessary, which are able to provide reliable identification and quantification of these compounds in biological matrices. Using charge-switch derivatization with AMPP ( N -(4-aminomethylphenyl)pyridinium chloride) in combination with liquid chromatography ion mobility quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS), we developed a non-target approach to analyze oxylipins in plasma, serum, and cells. The developed workflow makes use of an ion mobility resolved fragmentation to pinpoint derivatized molecules based on the cleavage of AMPP, which yields two specific fragment ions. This allows a reliable identification of known and unknown eicosanoids and other oxylipins. We characterized the workflow using 52 different oxylipins and investigated their fragmentation patterns and ion mobilities. Limits of detection ranged between 0.2 and 10.0 nM (1.0–50 pg on column), which is comparable with other state-of-the-art methods using LC triple quadrupole (QqQ) MS. Moreover, we applied this strategy to analyze oxylipins in different biologically relevant matrices, as cultured cells, human plasma, and serum. Graphical abstract Electronic supplementary material The online version of this article (10.1007/s00216-020-02795-2) contains supplementary material, which is available to authorized users.

Screening and Identification of Epoxy/Dihydroxy-Oxylipins by Chemical Labeling-Assisted Ultrahigh-Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry

“…Epoxy/dihydroxy-oxylipins are usually analyzed using LC-MS in a negative-ion mode. However, acidified eluents are commonly used for the separation of epoxy/dihydroxy-oxylipins by LC, which limits the generation of negatively charged ions, ultimately reducing the MS detection sensitivity of the analytes . Thus, DMED was used for derivatizing epoxy/dihydroxy-oxylipins to introduce tertiary amine groups into the analytes (Figure ).…”

“…A chemical derivatization-assisted LC-MS strategy with high sensitivity has been widely employed in the analysis of metabolites in low abundance. − However, few analytical methods based on this strategy for the profiling of oxylipins have been reported. − Hellhake et al used AMPP ( N -(4-aminomethylphenyl)­pyridinium chloride) to label oxylipins and developed a liquid chromatography-ion mobility-quadrupole time-of-flight mass spectrometry (LC-IM-QTOF-MS) for profiling oxylipins in biological samples . This reported method is able to improve the detection sensitivity of oxylipins, but it is still difficult to identify the structure of potential oxylipins by this method.…”

High Glucose Shifts the Oxylipin Profiles in the Astrocytes towards Pro-Inflammatory States

“…For example, the oxylipins of the n-6 group include both pro-inflammatory compounds and resolution substances, which are responsible for restoring the system after the pro-inflammatory stimulus has been applied [ 2 , 3 , 10 ]. The complexity of the oxylipin metabolism system has led to the development of mass spectrometric detection methods, making it possible to simultaneously measure several tens of oxylipins, derivatives of various PUFAs [ 11 , 12 , 13 ]. The measurement of oxylipin profiles reveals new steps in understanding the role of oxylipins in various pathologies.…”

Derivatization procedures and their analytical performances for HPLC determination in bioanalysis

“…time‐of‐flight mass spectrometry TOF–MS or ion trap mass spectrometry IT–MS) also became a useful technique for qualitative analysis in this domain (Chen, Wu, Tan, Zhu, & Chai, 2011; Williamson & Bartlett, 2007). Other advances in mass spectrometry, such as ion‐mobility spectrometry–mass spectrometry, also represent powerful tools in bioanalysis (Hellhake et al, 2020; Isenberg, Armistead, & Glish, 2014). Weak ionization, poor HPLC retention and instability of target analytes are some of the difficulties in the development of the HPLC–MS/MS method.…”

“…Other advances in mass spectrometry, such as ion-mobility spectrometry-mass spectrometry, also represent powerful tools in bioanalysis (Hellhake et al, 2020;Isenberg, Armistead, & Glish, 2014).…”